An orange (Citrus sinensis) is the most common fruits in the world. The wastes generated from the orange fruit needs to be put in to beneficial use. In this study the primary wastes (peel) of orange is use for preparation of prunin. a-L-Rhamnosidase (EC 3.2.1.40) secreted by Aspergillus flavipus MTCC-4644 are potential catalysis in hydrolysis of naringin content present in orange peels. a-L-rhamnosidase from the culture filtrate of a fungal strain, Aspergillus flavipus MTCC-4446 has been purified to homogeneity. The procedure involved concentration by ultra filtration and cation-exchange chromatography on carboxymethyl cellulose. The purified enzyme gave a single protein band corresponding to molecular mass of 40.0 kDa in SDS-PAGE analysis showing that the enzyme preparation was pure. The native PAGE analysis of the purified enzyme also gave single protein band confirming the purity of the enzyme preparation. Using p-nitro phenyl -a-L-rhamnopyranoside as substrate, Km and kcat values of the enzyme were 0.48 mM and 28. 4 s-1 respectively. The pH and temperature optima of the enzyme were 5.0 and 50 °C, respectively. The enzyme is stable below10ºC and at pH 4.5. The energy of activation for thermal denaturation of enzyme determined by Arrhenius plot was 32.06 k J mol-1.The enzyme hydrolyzed naringin content of orange peel to L-rhamnose and prunin.
Citrus sinansis, Naringin, a-L-Rhamnosidase, Prunin, Aspergillus flavipus, L- rhamnose
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